Dental practitioner should be able to detect and identify various oral lesions, hence provide accurate diagnosis and treatment plan to the patient for better prognosis. Some oral lesions which give similar clinical appearance must be confirmed by adjunctive procedures such as biopsy. They should be knowledgeable about various techniques used for oral biopsy and should have ability to understand and formulate treatment plan based on biopsy report. Thus the aim of the present study was to carry out a questionnaire based survey of dental practitioners in Madurai to assess their knowledge, practice  and attitude regarding biopsy procedures. This cross sectional questionnaire based study was done on 180 dental practitioners in Madurai,  consisting of 17questions formulated to collect information through online platform  on various aspects of oral biopsy. The data thus collected was tabulated. The response rate was 90%. Most of DPs had a good knowledge about biopsy but there were conflicting results on performing biopsy . Most of the dental surgeon are not comfortable performing biopsy with majority seem to lack experience and skill. There is need to organize training to Dental Surgeons to enhance their practical skills.